PRODUCT DATA SHEET
IncuCyte® Reagents
IncuCyte® Caspase-3/7 Reagent for Apoptosis
Catalog number: 4440
Presentation, storage and stability
The IncuCyte® Caspase-3/7 Reagent is supplied as one vial (20
µl) of 5 mM solution in dimethylsulfoxide (DMSO), with each
vial providing sufficient quantity for performing 100-200 tests
(1 test = 1 well of 96-well microtiter plate). Upon receipt, the
solution should be stored at 4°C. When stored as described the
stock solutions will be stable for at least 6 months.
Background and intended use
The IncuCyte Caspase-3/7 Reagent for Apoptosis couples
the activated caspase-3/7 recognition motif (DEVD) to a DNA
intercalating dye and is ideally suited to the mix-and-read, realtime quantification of cells undergoing caspase-3/7 mediated
apoptosis. Addition of the IncuCyte Caspase-3/7 Reagent
to normal healthy cells is non-perturbing to cell growth and
morphology. When added to tissue culture medium, the inert,
non-fluorescent substrate crosses the cell membrane where it is
cleaved by activated caspase-3/7 resulting in the release of the
DNA dye and green fluorescent staining of the nuclear DNA. With
the IncuCyte® integrated analysis software, fluorescent objects
can be quantified and background fluorescence minimized.
This reagent has been validated for use with the IncuCyte® livecell analysis system and enables the real-time evaluation of cell
apoptosis induced by pharmacological agents and/or genetic and
environmental factors. Furthermore, our IncuCyteCaspase-3/7
Reagent for Apoptosis can be combined with the IncuCyte®
system confluence metrics (phase), our range of IncuCyte®
NucLight nuclear labeling reagents, or the IncuCyte® Cytotox Red
Reagent to quantify cell proliferation or cytotoxicity alongside
apoptosis in a single well.
Recommended use
We recommend that IncuCyte Caspase-3/7 Reagent be used at
a final concentration of 5 µM (1:1000) in growth media and
added directly to cells in culture. When used in an IncuCyte®
live-cell analysis system, we recommend data collection every
2-3 hours.
Please see the relevant protocol published on our website:
essenbioscience.com/apoptosis
Safety data sheet (SDS) information
Download the SDS from our website: essenbioscience.com/apoptosis
Figure 1. IncuCyte Caspase-3/7 Reagent
Excitation and emission spectra of enzymatically-cleaved IncuCyte Caspase-3/7 reagent in the presence of dsDNA.
Absorption/emission maxima of IncuCyte Caspase-3/7 reagent bound to DNA: 500/530 nm.
IncuCyte.com
Page 1 of 2
PRODUCT DATA SHEET
IncuCyte® Caspase-3/7 Reagent for Apoptosis
Quick guide
1
2
SEED CELLS
Seed cells (100 µL/well)
into a 96-well plate.
PREPARE APOPTOSIS
REAGENT AND TREAT CELLS
3
Prepare the desired treatments at
1x in medium containing IncuCyte
Caspase-3/7 or Annexin V Reagents
and add treatment.
LIVE CELL FLUORESCENT
ANALYSIS
Capture images every 2-3
hours (20x or 10x) in the
IncuCyte® System. Analyze
using integrated software.
Figure 2. Concentration dependent activation of the IncuCyte Caspase-3/7 Reagent
by the DNA topoisomerase I inhibitor camptothecin in HT-1080 fibrosarcoma cells.
B
A
(A) Time course of apoptosis in the presence of increasing concentrations
of camptothecin. Apoptosis has been quantified as the number of green
fluorescent caspase-3/7 active objects for each time point.
(B) Concentration response curve to camptothecin. Area under
the curve (AUC) values have been determined for the time course
shown in panel A.
FOR RESEARCH USE ONLY. NOT FOR THERAPEUTIC OR DIAGNOSTIC USE.
Product
IncuCyte® Caspase-3/7 Reagent for Apoptosis
Cat No.
Amount
Ex. maxima
Em. maxima
4440
20 µl
500 nm
530 nm
Product label licence
This product is provided under an Agreement between Biotium and Essen BioScience. The manufacture, use, sale, offer for sale, or import of this product is subject to one or
more patents or pending applications owned or licensed by Biotium, Inc. (U.S. patent application No. US60/741,263). The purchase price of this product includes a limited,
non-transferable immunity from suit under U.S. Patent application No. US60/741,263 and corresponding patent claims outside of the United Sates, to use only the purchased
amount of the product and components of the product solely for the buyer’s own internal research (whether the buyer is an academic or for-profit entity), in a manner
consistent with the accompanying product literature. No right to use, sell or otherwise transfer (a) this product (b) its components or (c) materials made using this product or
its components to a third party, or otherwise use this product or its components or materials made using this product or its components for any other purpose or a Commercial
Purpose, is hereby granted expressly, by implicate or by estoppel. Commercial Purpose mean any activity by a party for consideration and include, but is not limited to : (1)
use of the product or its components in manufacturing; (2) use of the product or its components to provide a service including without limitation, reporting information or
data resulting from use of the product or its components for a fee or other commercial consideration; (3) use of the product or its component for therapeutic, diagnostic or
prophylactic purposes; or (4) resale of the product or its components, whether or not such product or its components are resold for use in research. For products that are subject
to multiple limited use label licenses, the most restrictive terms apply. This product is for research use only. Diagnostic uses require a separate license from Biotium, Inc. For
information on purchasing a license to this product including for purposes other than research, contact Biotium, Inc., 3159 Corporate Place, Hayward, CA 94545, Tel: (510)
265-1027. Fax: (510) 265-1352. Email: [email protected]. All names containing the designation are trademarks of Biotium, Inc.
IncuCyte.com
Essen BioScience, 300 West Morgan Road, Ann Arbor, Michigan, USA 48108
© 2016 Essen BioScience. All rights reserved. IncuCyte®, Essen BioScience® and all names of Essen BioScience
products are registered trademarks and the property of Essen BioScience unless otherwise specified.
Page 2 of 2
8000-0069-G00